EXPERIMENT NUMBER: ONE

Degree of dissociation of double salts and complex compounds

1. Objective
The objective of the experiment is to:
 to identify degree of dissociation of double salts and complex compounds
2. Theory:
Double salts are salts containing more than one cation or anion, and are obtained by combination of two
different salts which were crystallized in the same regular ionic lattice.
Note that double salts should not be confused with a complex. When double salts dissolved in water, it
completely dissociates into simple ions while complexes do not; the complex ion remains unchanged. For
example, KCeF4 is a double salt and gives K+, Ce3+ and F− ions when dissolved in water, whereas
K4[YbI6] is a complex salt and contains the discrete [YbI6]4− ion which remains intact in aqueous
solutions. It is therefore important to indicate the complex ion by adding square brackets "[ ]" around it.
Double salts loose their identity in aqueous solution or any other solvent.
Eg Mohr’s salt ((NH4)2SO4.FeSO4.6H2O), Nessler’s reagent (K2[HgI4], Chrome alum
(K2SO4.Cr2(SO4)3.24H2O), Alum (KAl(SO4)2·12H2O), etc.

The difference between double salts and complexes is shown below.

Double Salt Complex Compound

Bond present in it is electrovalent and Bond present in it is electrovalent, covalent and
covalent coordinate covalent
They exist in solid state They exist in solid state as well as in aqueous solutions
They lose their identity in aqueous solution They do not lose their identity in aqueous solution
They contain two salts in equimolar ratio They contain ions which may or may not be in
equimolar ratio
The metal ions show their normal valencies The metal ions show primary and secondary valencies
They form simple ions in aqueous solution They form complex ions in aqueous solution
They are less stable They are more stable

Examples of Double salts:

KCl  MgCl2  6 H 2 O  KCl..MgCl2 .6 H 2 O (Carnollite)
K 2 SO4  Al2 ( SO4 ) 3  24 H 2 O  K 2 SO4 . Al 2 ( SO4 ) 3 .24 H 2 O (Potash alum)
Examples of Complexes:
Fe(CN ) 2  4 KCN  K 4 [ Fe(CN ) 6 ]
CuSO4  4 NH 3  [Cu ( NH 3 ) 4 ]SO4

3. Apparatus and Chemical

A. Apparatus: Test tube, holder, spatula, tripod-Gauze
B. Chemicals: Mohr’s salt, Nessler’s reagent, water, ammonium sulfide, NaOH, BaCl2, Conc.NH3,
Potassium Ferrocyanide, Hydrous copper sulfate, dilute HCl, AgNO3, FeCl3

4. Procedure
Ferrous Ammonium Sulphate
 Take Mohr’s salt ((NH4)2SO4.FeSO4.6H2O), Dissolve it in distilled water and test for the
individual ions
2+
Fe - add K4[Fe(CN)6] solution, Note any change
NH4+ - Add NaOH solution and heat, Note any change.
What type of gas is evolved? Use litmus paper and check the type of gas evolved.
SO42- - Add BaCl2 solution, and check the solubility of precipitate in dilute HCl.
Hydrous Copper Sulphate
 Dissolve the salt in distilled water and add conc. Ammonia Solution
Q. What have you seen in both steps, and why?
Potassium Ferrocyanide
Dissolve some in distilled water and perform the following tests.
Fe2+ - Test with Ammonium Sulphide solution. (Don’t add excess!)
CN- - Add silver nitrate solution. (Don’t add excess!)
[Fe(CN)6]4- - Add ferric chloride solution , and observe any change.
Add aqueous sodium hydroxide to the solid, or solution, under test and warm the mixture. Result: If
ammonium ions are present then a pungent-smelling gas is produced. The gas produced turns damp red
litmus paper blue. It is ammonia, NH3.

Testing for the sulfate ion. You can test to see if a solution contains sulfate ions by using barium
chloride. If barium chloride solution is added to a sample of water containing sulfate ions, barium sulfate
is formed. Barium sulfate is insoluble in water, and will be seen as a white precipitate.

"Make a solution of the given salt (whose cation is to identified) and add potassium ferrocyanide [
potassium hexacyanidoferrate(II) ] solution to it.

If a dark blue precipitate is formed, then the presence of ferrous ions in the given salt is confirmed."
 Experiment 2
Preparation of Ferrous Ammonium Sulphate or Ammonium Iron (II) Sulfate Hexahydrate

Aim: To prepare double salt

Theory

Iron is a d-block element and a transition metal. Like most d-block elements, it exhibits various
oxidation states. Compounds are known that contain iron in all of the oxidation states from 0 to
+7. Iron reacts with acids that have non-oxidizing anions, such as hydrochloric acid or sulfuric
acid, evolving hydrogen gas and forming iron(II) salts. If iron is dissolved in an acid that has an
oxidizing anion, the iron(III) salt is obtained. Thus when iron is dissolved in nitric acid, it is
oxidized to iron(III) and the product is iron(III) nitrate. At the same time, the nitric acid is
reduced to various oxides of nitrogen including NO2 (brown fumes).
The iron(II) ion is not very stable under neutral conditions and is slowly oxidized to iron(III)ion
(and precipitates as the hydroxide). Aqueous solutions of iron(II) ions are more stable in acidic
solution, so the aqueous iron(II) sulfate that is provided in the laboratory is acidified with
sulfuric acid.
The ferrous ammonium sulfate is also known as Mohr's salt, used to provide a stable source of ferrous
ions for oxidation-reduction titrations. Mohr’s salt (Ferrous Ammonium Sulphate) is inorganic
compound with the formula (NH4)2Fe(SO4)2.6H2O containing two different cations, Fe2+ and NH4+,
it is classified as a double salt of ferrous sulfate and ammonium sulfate.
When a mixture containing equimolar proportions of ferrous sulphate and ammonium sulphate is
crystallized from its solution, a double salt is formed. The formation of double salt may be shown as
follows:
FeSO4  ( NH 4 ) 2 SO4  6 H 2 O  ( NH 4 ) 2 SO4 .FeSO4 .6 H 2 O

Mohr salt is a common laboratory reagent. Like the other ferrous sulfate salts, ferrous ammonium sulfate
dissolves in water to give the aquo complex [Fe(H2O)6]2+, which has octahedral molecular geometry.
Ferrous sulphate is mainly used as precursor to other iron compound. It also used as a reducing
agent.
 Apparatus and Chemicals

Beaker (50 mL) Wire gauze

Glass rod Ferrous sulphate
Conical flask (50 mL) Ammonium sulphate
Tripod stand Dilute sulphuric acid
Funnel Ethanol

Procedure

 Dissolve 3.5 g of ferrous sulphate and 1.7 g of ammonium sulphate (weighed separately), in 5 mL
of distilled water contained in a 50 mL conical flask by heating. Add about 0.5 mL of dilute
sulphuric acid to the flask and concentrate the solution by heating till the crystallization point is
reached.
 Allow the mixture to cool to room temperature slowly.
 On cooling, light green crystals of ferrous ammonium sulphate separate out.
 Decant the mother liquor and wash the crystals by shaking with very small amounts of 1:1 cold
water and alcohol mixture to remove sticking mother liquor.
 Separate the crystals by filtration wash with alcohol, dry between the folds of a filter paper and
record the yield.
Result: Calculate the yield.

Precautions

 Cool the solution slowly to get good crystals. Avoid rapid cooling.
 Do not disturb the solution while cooling.
 Avoid prolonged heating while preparing crystals of ferrous ammonium sulphate, as it may oxidise
ferrous ions to ferric ions and change the stoichiometry of the crystals.
Questions

a. Why do we take equimolar quantities of reacting compounds in the preparation of double salts?
b. In the preparation of ferrous ammonium sulphate, can concentrated sulphuric acid be used in place
of dilute sulphuric acid? Explain.
c. What is the difference between iron compounds; K4[Fe(CN)6] and .(NH4)2(FeSO4)2.6H2O?
Tests (Prepare a solution from the compound prepared above)
Fe2+Add NaOH solution and note any change.
Add ammonium sulfide solution. What do you observe?
Add potassium hexacyanoferrate(III) solution, and note the color formed.
NH4+ Add NaOH solution, and heat it gently. What does happen to it?
Add Nessler’s reagent and see any change.
Add sodium cobalt nitrite solution. Is there any change?
SO42-adds BaCl2 solution, and check the solubility of the precipitate in dilute HCl.

OR
It can prepared from iron metal.
Chemicals Required
Ammonium sulfate 6.0 g
Iron 6.0 g
2.0 mol / L sulfuric acid 50.0 mL
Procedure
In an 250 mL Erlenmeyer flask, dissolve the ammonium sulfate, (NH4)2SO4, in 12 mL of deionized
water. Add the iron to a second 250 mL Erlenmeyer flask with the sulfuric acid. Fit the flask
containing the solid iron with a cotton wool or glass wool plug. Using a hot plate, heat the flask
gently until about 2 cm of foam forms on the surface of the liquid. Maintain this rate of reaction
for about 20 minutes by adjusting the heat as required. This may mean periodically removing the
flask from the hot plate. Do NOT leave your bench until the reaction is complete.
A little iron may remain. The flask may be swirled a few times (care!!) during the reaction but too
much agitation will lead to excess evaporation of water and precipitation of the intermediate
product (FeSO4).
Filter the solution directly into the flask containing the ammonium sulfate solution. Discard the
unreacted iron. Mix the solution by gentle swirling and then allow it to cool first to room tem-
perature and then in an ice bath. After a minimum of 30 minutes of cooling in the ice bath, swirl
the mixture vigorously and then return it to the ice bath for a further 10 minutes.
Collect the crystalline precipitate by suction filtration using a Buchner funnel. Do NOT wash the
crystals with water! Wash the crystals once with 10 mL of 50% methanol (prepare this yourself: 5
mL of methanol mixed with 5 mL of deionized water) and then once with 10 mL of methanol. Air
dry on a watch glass in your locker for a week. Determine the mass of the dry product.

6
Questions
1. What is a primary standard? When could your product be used as a primary standard.
Include a balanced reaction equation for the appropriate reaction.
2. Define double salt and complex ion. Give an example of each and explain how they differ.
References
1. G. Svehla. Vogel's Qualitative Inorganic Chemistry, sixth edition.
2. Petrucci, Harwood et al., General Chemistry, 9th or 10th edition.
3. D.C. Harris, Quantitative Chemical Analysis, any edition.

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Experiment 3: Preparation and Analysis of Potassium cis-diaquadioxalatochromate (III)
dehydrate

Theory: Potassium cis-diaquadioxalato chromate ( III) dihydrate, K[Cr(H2O)2 (C2O4)2].2H2O, is

prepared from oxalic acid dihydrate, potassium dichrom a te, carbon dioxide and water.
The reaction equation is:

Potassium cis-diaquadioxalatochromate (III) dehydrate may conduct electricity when it dissolved

in water due to the presence ions in the solution.
Molar conductivity: is defined as the conductivity of an electrolyte solution divided by the molar
concentration of the electrolyte, and so measures the efficiency with which a given electrolyte conducts
electricity in solution. It is the conducting power of all the ions produced by dissolving one mole of an
electrolyte in solution. Its units are siemens per meter per molarity.
The usual symbol is a capital lambda, Λ, or Λm.
Friedrich Kohlrausch's researches in 1875-79 established that to a high accuracy in dilute solutions,
molar conductivity is composed of individual contributions of ions. This is known as the law of
independent migration of ions.
From its definition, the molar conductivity is given by:
Λm =k/c, where κ is the measured conductivity (formerly known as specific conductance) and c is the
electrolyte concentration.
Two cases should be distinguished: strong electrolytes and weak electrolytes.
For strong electrolytes, such as salts, strong acids and strong bases, the molar conductivity depends only
weakly on concentration.
For weak electrolytes (i.e. incompletely dissociated electrolytes), however, the molar conductivity
strongly depends on concentration: The more dilute a solution, the greater its molar conductivity, due to
increased ionic dissociation. For example, acetic acid has a higher molar conductivity in dilute aqueous
acetic acid than in concentrated acetic acid.

8
Procedure: A mixture of 0.5g of Potassium dichromate and 1.5g of oxalic acid dihydrate is ground
together thoroughly in a dry mortar. The mixture is placed in a compact pile in the center of the
evaporating dish whose inner surface has been moistening with water. The dish is covered with a watch
glass and placed on a hot plate that is just hot to touch. A spontaneous reaction with vigorous
effervescence beings almost immediately and the mixture liquefies to syrup. Continue heating till thick
viscous slurry is obtained, then 2.5 ml of 95% ethanol is added and warming is continued. The mixture is
triturated first with spatula and then with pestle until the mixture solidifies. If solidification does not
occur, the ethanol is decanted. An additional 2.5 ml of ethanol is added and the trituration with warming
is continued until the product is granular. The black product is collected on a Buchner funnel, washed
with 1 ml portions 95% of ethanol, 0.5 ml of ether and then vacuum dried. Take the weight of the
complex and calculate the yield.
Note: Keep the sample in desiccators
INSTRUMENTATION ANALYSIS
1) Measure its specific conductance in distilled water and calculate the molar conductance.
VOLUMETRIC ANALYSIS
Analysis of the Oxalate content in K[cis-Cr(H2O)2(OX)2].2H2O using potassium permanganet
Weigh 0.3g of the dried K[cis-Cr(H2O)2(OX)2].2H2O in a beaker, and add 10mL water followed by 10
mL potassium hydroxide(10%). Heat the mixture until Cr(OH)2 (green precipitated) is completely
precipitated (about 10min). Filter off the precipitate and acidify the filtrate with 2N H2SO4 and add 10 ml
of excess of 2N H2SO4 and heat it to about 700C. Titrate against standard potassium permanganate
(0.1M). Calculate the concentration of the oxalic acid.

9
 Note: Standardization of KMnO4 should be done.

Experiment 8 – Redox Titrations

Potassium permanganate, KMnO4 is a strong oxidizing agent. Permanganate, MnO4-, is an intense dark
purple color. Reduction of purple permanganate ion to the colorless Mn+2 ion, the solution will turn
from dark purple to a faint pink color at the equivalence point. No additional indicator is needed for this
titration. The reduction of permanganate requires strong acidic conditions. In this experiment,
permanganate will be reduced by oxalate, C2O42- in acidic conditions. Oxalate reacts very slowly at
room temperature so the solutions are titrated hot to make the procedure practical. The unbalance redox
reaction is shown below.

In part I of this experiment, a potassium permanganate solution will be standardized against a sample o
potassium oxalate. Once the exact normality (eq/L) of the permanganate solution is determined, it can
be used as a standard oxidizing solution. In part II of this experiment, the standard permanganate
solution will be used to find the concentration of iron(II) in a ferrous solution (g/L). The unbalanced
redox reaction is shown below.

Phosphoric acid will be used to ensure that the ferric product, Fe3+ remains in its colorless form.

10
Part (I) - Preparation of a 0.1 N KMnO4 Solution.
1. On a centigram balance, weigh about 1.0 g KMnO4 crystals on a piece of weighing paper. Add the
crystals to a 500 mL Florence Flask.
2. Add about 350 mL of distilled water to the flask.
3. Heat the solution with occasional swirling to dissolve the KMnO4 crystals. Do not boil the solution.
This may take about 30 minutes.
4. Allow the solution to cool and stopper. You will need this solution for both day 1 and day 2.
Part (II) - Standardization of a KMnO4 solution.
1. On weighing paper, weigh about 0.2 – 0.3 g of K2C2O2H2O on the analytical balance. Record the
exact mass. Transfer the sample to a 250 mL Erlenmeyer flask.
2. Rinse and fill the buret with the KMnO4 solution.
3. Add 50 mL of distilled water and 20 mL of 6 N H2SO4 to the oxalate sample in the Erlenmeyer flask.
Swirl to dissolve the solids.
4. Heat the acidified oxalate solution to about 85 °C. Do not boil the solution.
5. Record the initial buret reading. Because the KMnO4 solution is strongly colored, the top of the
meniscus may be read instead of the bottom.
6. Titrate the hot oxalate solution with the KMnO4 solution until the appearance of a faint pink color.
7. Record the final buret reading and calculate the volume of KMnO4 used in the titration.
8. Discard the titration mixture down the drain and repeat the titration with a new sample of oxalate for a
total of 2 trials.
9. An oxalic acid solution may be used to wash the buret and the titration flask if a brown stain remains
in the glassware.

Calculations
1. Using the half-reaction method, write a balanced redox equation for the reaction of permanganate
with oxalate in an acidic solution.

11
2. Calculate the equivalent weight of the oxalate reducing agent from the molar mass of the oxalate
sample and the equivalence of electrons lost by the reducing agent in the oxidation half-reaction.

3. Use the sample mass and the equivalent weight to calculate the number of equivalents of oxalate in
each sample.

At the equivalence point, the equivalence of the reducing agent is equal to the equivalence of the
oxidizing agent.

4. Calculate the normality of the KMnO4 solution from the equivalence of the oxidizing agent and the
volume used in the titration.

12
 EXPERIMENT NUMBER: FOUR
Preparation of HexammineCobalt (III) chloride
Theory: HexammineCobalt (III) chloride is the chemical compound with the formula [Co(NH 3)6]Cl3.
This coordination compound is considered as ‘Werner complex’, name after the pioneer of coordination
chemistry, Alfred Werner. This salt consists of cations of [Co(NH3)6]3+ and three chloride ions. Originally
this complex considered as yellow complex.
Properties and Structure
 The chloride ions in [Co(NH3)6]Cl3 can be exchanged with variety of other anions such as
Bromide, Iodide, etc.
 Chemical formula: H18N6Cl3Co  Density: 1.71 g/cm3
 Molar mass: 267.48g/mol  Coordination geometry: octahedral
 Appearance: yellow/orange crystal
Preparation
 Cobaltammine chloride is prepared from cobalt (II) chloride.
 The later is treated with ammonia and ammonium chloride followed by oxidation.
 Oxidants include: H2O2/O2 in the presence of charcoal catalyst.
 Chemical equation:
2CoCl2.6H2O +2NH4Cl +10NH3 (aq) + H2O2 (aq) +3H2O(l) 2[Co(NH3)6]Cl3(s) +1/2O2(g)
Uses
[Co(NH3)6]3+ is a component of some structural biology method (specially for DNA or RNA where +ve
ion stabilize tertiary structure of the phosphate back bone.
Procedure:
1. Place 1gm of decolorizing charcoal in a conical flask.
2. Dissolve 9 gm of Cobalt (II) chloride and 6 gm of ammonium chloride in 10ml of distilled water with
heating.
3. The hot solution should be transferred to a conical flask containing the charcoal.
4. To this mixture add 20ml of Conc.NH3 solution.
5. Cool the mixture under cold tap water.
6. Add slowly 20ml of 20% H2O2 very slowly with continuous stirring.
7. After adding all H2O2 heat the mixture to 60oC and maintain this temperature for 15 to 20 minutes
until the pink color of the solution is disappeared. (Be careful while heating)
8. Cool the reaction mixture in cold tap water first, and then in an ice cold water.
9. Filter the solid in the Buchner funnel and transfer the precipitate to a beaker containing a boiling
solution of 3ml Conc.HCl and 80ml water. When the entire solid, except the charcoal, has dissolved
filter the hot suspension.
10. Add 10ml Conc. HCl to the filtrate and cool the solution in ice bath.
11. Filter the crystals on a Buchner funnel and dry between filter papers. Record your yield and calculate
the percentage yield.

13
Experiment 5: Preparation of potassium bis(oxalato)cuprate(II) dihydrate,
K[Cu(C2O4)2].2H2O

UV-Visible Spectroscopy
Absorption Measurements & their Application to Quantitative Analysis

A study of the interaction of light (or other electromagnetic radiation) with matter is an important and
versatile tool for the chemist. Indeed, much of our knowledge of chemical substances comes from their
specific absorption or emission of light. In this experiment, we are interested in analytical procedures
based on the amount of light absorbed (or transmitted) as it passes through a sample.
Suppose you look at two solutions of the same substance, one a deeper color than the other. Your
common sense tells you that the darker colored one is the more concentrated. In other words, as the color
of the solution deepens, you infer that its concentration also increases. This is an underlying principle of
spectrophotometry: the intensity of color is a measure of the amount of a material in solution.
A second principle of spectrophotometry is that every substance absorbs or transmits certain wavelengths
of radiant energy but not other wavelengths. For example, chlorophyll always absorbs red and violet light,
while it transmits yellow, green, and blue wavelengths. The transmitted and reflected wavelengths appear
green the color your eye “sees.” The light energy absorbed or transmitted must match exactly the energy
required to cause an electronic transition (a movement of an electron from one quantum level to another)
in the substance under consideration. Only certain wavelength photons satisfy this energy condition.
Thus, the absorption or transmission of specific wavelengths is characteristic for a substance, and a
spectral analysis serves as a “fingerprint” of the compound.

In recent years spectrophotometric methods have become the most frequently used and important
methods of quantitative analysis. They are applicable to many industrial and clinical problems involving
the quantitative determination of compounds that are colored or that react to form a colored product.

LIGHT AND THE PERCEPTION OF COLOR

Light is a form of electromagnetic radiation. When it falls on a substance, three things can happen:
o the light can be reflected by the substance
o it can be absorbed by the substance
o certain wavelengths can be absorbed and the remainder transmitted or reflected

14
Since reflection of light is of minimal interest in spectrophotometer, we will ignore it and turn to the
absorbance and transmittance of light.
The color we see in a sample of solution is due to the selective absorption of certain wavelengths of
visible light and transmittance of the remaining wavelengths. If a sample absorbs all wavelengths in the
visible region of the spectrum, it will appear black; if it absorbs none of them, it will appear white
or colorless. We see the various colors when particular wavelengths of radiant energy strike our eyes. For
example, the wavelength we perceive as green is 0.0000195 inches or, expressed more scientifically, 495
nanometers.
Suppose we shine a beam of white light at a substance that absorbs blue light. Since the blue component
of the white light gets absorbed by the substance, the light that is transmitted is mostly yellow, the
complementary color of blue. This yellow light reaches our eyes, and we “see” the substance as a yellow
colored substance. The table below gives pairs of complementary colors and the corresponding
wavelength ranges.

You should remember, of course, that the visible range is only a very small part of the electromagnetic
spectrum. Ultraviolet and infrared spectrophotometric methods are suitable for many colorless substances
that absorb strongly in the UV or IR spectral regions.

TRANSMITTANCE, ABSORBANCE, AND THE BEER-LAMBERT LAW

We define transmittance as the ratio of the amount of light transmitted to the amount of light that initially
fell on the surface.

15
Absorbance is defined as the negative logarithm of the transmittance, and you will note that absorbance
and transmittance bear an inverse relationship.
Absorbance = - log T = - log P/Po

Going back to our example of chlorophyll, if you have two colored solutions, you may deduce that the
darker colored green solution appears darker because it absorbs more of the light falling on it. Because the
darker solution is also the more concentrated one, you can also say that the more concentrated one
absorbs more of the light. That is, the absorbance increases as concentration increases.
Next, suppose that there are two test tubes, both containing the same solution at the same concentration.
The only difference is that one of the test tubes is thicker than the other.

We shine light of the same intensity (Po) on both containers. In the first case the light has to travel
through only a short distance, whereas in the second case it has to pass through a much longer length of
the sample. We might deduce that in the second case more of the light will be absorbed or cut off,
since the path length is longer. In other words, absorbance increases as path-length increases.
The two observations described above (those dealing with the relationship between absorbance and
concentration and absorbance and path length) constitute the BEER-LAMBERT LAW.
Beer-Lambert Law
Absorbance ∝ path length ( l ) • concentration (c)
A= εlc where
• A is a dimensionless number.
• ε the proportionality constant, is called the molar extinction coefficient or molar absorptivity. It is a
constant for a given substance, provided the temperature and wavelength are constant. It has units of
liter/mol.cm. l and c have the usual units of length (cm) and concentration (mol/liter).

The quantitative measurement of light absorption as a function of wavelength can establish both the
identity and the concentration of a substance in solution.
The spectrophotometer is an instrument that separates electromagnetic radiation into its component
wavelengths and selectively measures the intensity of radiation after passing through a sample.

16
In conventional spectrometers electromagnetic radiation is passed through the sample which is held in a
small square-section cell (usually 1 cm wide internally) called Cuvette. There are three types of cuvette.
These are plastic glass and quartz. A common feature of all cuvette is that each one has 2 opposite opaque
sides and 2 opposite transparent sides. You will hold the cuvette by the 2 opaque sides. The transparent
sides are meant for the pathway of UV/Visible light.
Radiation across the whole of the ultraviolet/visible range is scanned over a period of approximately 30s,
and radiation of the same frequency and intensity is simultaneously passed through a reference cell
containing only the solvent. Photocells then detect the radiation transmitted and the spectrometer records
the absorption by comparing the difference between the intensity of the radiation passing through the
sample and the reference cells (Fig. 1).

Figure 1: Diagram showing how the ultraviolet/visible spectrometer works

No single lamp provides radiation across the whole of the range required, so two are used. A hydrogen or
deuterium discharge lamp covers the ultraviolet range, and a tungsten filament (usually a
tungsten/halogen lamp) covers the visible range. The radiation is separated according to its
frequency/wavelength by a diffraction grating followed by a narrow slit. The slit ensures that the radiation
is of a very narrow waveband. i.e it is monochromatic.
Detection of the radiation passing through the sample or reference cell can be achieved by either a
photomultiplier or a photodiode, that converts photons of radiation into tiny electrical currents; or a
semiconducting cell (that emits electrons when radiation is incident on it) followed by an electron
multiplier similar to those used in mass spectrometers. The spectrum is produced by comparing the
currents generated by the sample and the reference beams.

17
 SPECTROPHOTOMETRIC ANALYSIS
This section of the introductory material outlines a general approach for spectrophotometric analysis, first
finding the absorption spectrum of “finger prints” of a substance and then determining its concentration.
1. Plotting Absorption Spectra
Recall that the extinction coefficient for any given substance is a constant only so long as the wavelength
of light is constant. You will see that the absorbance changes with wavelength. The plot of a sample's
absorbance of light at various wavelengths is called its absorption spectrum. (The abscissa or horizontal
axis may be expressed in terms of wavelength and the ordinate or vertical axis in terms of absorbance.)
The plot below gives the absorption spectrum of potassium permanganate (KMnO4), a purple colored
solution, at two different concentrations. Curves 1 and 2 represent the absorption spectra measured under
the same conditions except that curve 1 represents a more concentrated solution than curve 2.
Note the similar shapes of the curves.
2. Choice of Wavelength
According to the Beer-Lambert Law absorbance is proportional to concentration at each wavelength.
Theoretically we could choose any wavelength for quantitative estimations of concentration. However,
the magnitude of the absorbance is important, especially when you are trying to detect very small
amounts of material. In the spectra above note that the distance between curves 1 and 2 is at a maximum
at 525 nm, and at this wavelength the change in absorbance is greatest for a given change in
concentration. That is, the measurement of concentration as a function of concentration is most sensitive
at this wavelength. For this reason we generally select the wavelength of maximum absorbance for a
given sample and use it in our absorbance measurements.

18
Suppose instead that we had chosen a wavelength of 500 nm for our measurement, this wavelength being
on one of the steep portions of the curve. Examination of the curve shows that even a small fluctuation in
the wavelength will cause a large error in the absorbance. Most spectrophotometers show a slight
fluctuation in the wavelength, so errors in absorbance will be magnified if we select a wavelength such as
500 nm in our preceding example.
3. Plotting Calibration Graphs
Once we have chosen the correct wavelength, the next step is to construct a calibration curve or
calibration plot. This consists of a plot of absorbance versus concentration for a series of standard
solutions whose concentrations are accurately known. Because calibration curves are used in reading off
the unknown concentrations, their accuracy is of absolute importance. Therefore, make the standard
solutions as accurately as possible and measure their absorbance carefully. Each standard solution should
be prepared in identically the same fashion, the only difference between them being their concentrations.
When drawing the calibration graphs, take care not to lose any of the accuracy of the experimental data
by choosing axes that are too small. Choose axes to represent the accuracy possible in reading the
instrument. For example, if it is possible to read absorbance correct to the second decimal place, say 0.47,
then construct the absorbance axis so that 0.47 can be located accurately on it.

Slope of the best straight line through the data points in the calibration plot is 1.65. Plot intercept is 0.008.

Equation of straight line:

Absorbance = 1.65 (Concentration) + 0.008
To find an unknown concentration for a sample, subtract the intercept from the absorbance reading and
divide the result by the slope. Here the equation would be

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The Beer-Lambert Law (A = ε l c) implies that when concentration is equal to zero (c = 0), absorbance
must also be zero (A = 0). In other words, the calibration line must pass through the origin.
A major source of error in spectrophotometric analysis is applying the Beer-Lambert Law at
inappropriate concentrations. The Beer-Lambert Law is strictly applicable only for dilute solutions. It
becomes less and less accurate as the concentration of the solution increases.
Once you have the calibration curve set up, you can measure the absorbance of any unknown solution at
the same wavelength and read off its concentration from the graph or calculate from the slope.

Spectroscopy of Copper complexes

Uncomplexed copper(II) ions appear white or colourless because the transition between the highest
occupied orbital and the lowest unoccupied orbital is at too short a wavelength to see. In hydrated copper
(II) ions the energy levels of the 3d orbitals are split (Fig. 3), and when visible light is absorbed a
transition is possible. For the hydrated copper (II) ion the absorption occurs at the red end of the spectrum
( Fig. 4) hence the complex appears blue. The amount of splitting of the energy levels depends on the
ligand, so if ammonia replaces water and ∆E increases, the colour of the complex becomes blue/violet i.e
absorption occurs at the middle of the visible spectrum (Fig. 5) . Note that ε is larger (both spectra
contain the same concentration of copper ions).

Figure 3: Splitting of d orbital energy levels in Cu(H 2O)62+

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Figure 4 Ultraviolet/visible spectrum of Cu(H2O)62+

Figure 5: Ultraviolet/visible spectrum of Cu(NH3)4(H2O)22+

Changing the ligands around a transition metal ion can increase the stability of the complex formed
because the difference between the energy levels of the d orbitals increases ( Fig. 6 ). (Stability in this
context is taken to be the extent to which the complex will form from, or dissociate into, its constituents at
equilibrium.)

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Figure 6: Increased splitting of d orbital energy levels

Some ligands giving increasing separation of energy levels (and therefore increasing stability to the ions)
are:

This list forms part of the spectrochemical series and the order can vary depending on the metal ion in the
complex.
Stability of complex ions
It is possible to measure the relative stabilities of complexes through equilibrium constants called stability
constants. If the successive replacement of water molecules by ammonia molecules in the Cu(H2O)62+ ion
is used as an example, the first substitution can be described by the equation:

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Complexes with large stability constants are more stable than those with small constants. For example,
using chloride ions to replace the water ligands surrounding copper ions gives a less stable complex. This
is demonstrated simply by adding aqueous ammonia to a solution containing the CuCl42– ion – the
solution turns from yellow to deep blue. The reduced stability is also evident in the smaller stability
constants:
K1 = 631, K2= 398, K3= 3.09, K4= 5.37, K = 4.17 x 105

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Procedure: Dissolve 1g potassium oxalate monohydrate (K2C2O4.H2O) in 2.7 mL distilled water
and heat the solution to 90oC. Dissolve 0.3 g copper sulphate penthydrate (CuSO4.5H2O) in
0.6mL distilled water, and heat the solution to 90oC. Filter the solution while still hot, and slowly
with stirring, add the hot filtrate to the hot solution of potassium oxalate. Cool the mixture in an ice bath,
filter the crystals formed and wash with cold water, followed by ethanol and finally acetone. Dry the
crystals in air. Record your yield.

INSTRUMENTATION ANALYSIS
1) Measure its specific conductance in distilled water and calculate the molar conductance.
2) Record its visible spectrum in distilled water and compare its λmax.

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